Extraction method of soil protein

An extraction method and protein technology, which is applied in the field of soil protein extraction, can solve the problems of lack of soil protein extraction methods and the ineffective development of soil proteomics, and achieve the effects of eliminating humic acid interference, preventing degradation, and reducing impact

Inactive Publication Date: 2017-05-10
NORTHEAST AGRICULTURAL UNIVERSITY
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Problems solved by technology

The extraction of soil protein is the premise of soil proteomics research. However, due to the lack of soil protein extraction methods, the current soil proteomics research cannot be carried out effectively.

Method used

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  • Extraction method of soil protein
  • Extraction method of soil protein

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Experimental program
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Embodiment

[0055] 1 Soil protein extraction steps:

[0056] Extraction of soil protein: Weigh 2g of terrestrial soil, 2g of tomato rhizosphere soil, and 2g of tomato rhizosphere soil after returning wheat straw to the field, put the three kinds of soil into 10ml centrifuge tubes, add 5ml of soil protein extraction buffer, and the maximum Shake at a speed of 15 minutes, let stand for 3 minutes, open the cover of the centrifuge tube, heat the centrifuge tube in boiling water for 10 minutes, take out the centrifuge tube and place it in ice water at 0°C, cool for 3 minutes, shake at the maximum speed for 5 minutes, centrifuge at 5000 rpm at 4°C for 5 minutes, and remove the supernatant Pass the liquid through a 0.45μm nylon filter membrane, transfer the supernatant of the filtered membrane to a new 10ml centrifuge tube, add 2ml of Tris saturated phenol with pH 8.0, shake at the maximum speed for 20min, then centrifuge at 12000rpm at 4°C for 20min, remove the supernatant , remove the lower ph...

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Abstract

The invention provides an extraction method of soil protein. The extraction method comprises the following steps of adding soil protein extraction buffer liquid into a soil sample, oscillating, and standing; putting into boiling water for heating, cooling in 0 DEG C ice water, oscillating, and centrifuging; filtering supernatant, adding Tris saturated phenol into the filtered supernatant, oscillating, centrifuging, removing the supernatant, and collecting lower phenol solution; adding an ammonium acetate methanol solution into the lower phenol solution, and storing for 6 to 8h at the temperature of -20 DEG C; centrifuging, removing out the supernatant, collecting protein precipitate, cleaning the protein precipitate by acetone, and freeze-drying, so as to obtain protein dry powder. After detecting by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gelelectrophoresis) electrophoresis, compared with the protein band obtained by other two representative extraction methods of the soil protein, the band of the soil protein extracted by the method is clearer.

Description

technical field [0001] The invention relates to a method for extracting soil protein. Background technique [0002] Environmental metaproteomics is a research technique that has emerged in recent years. This technique can effectively analyze the complex microbial populations in soil to reveal changes in soil ecosystems. [0003] The key to the study of metaproteomics in the soil environment is the extraction and analysis of soil proteins, and the extraction of proteins is the focus of the study of the soil environment. The extraction of soil protein is the premise of soil proteomics research. However, due to the lack of soil protein extraction methods, the current soil proteomics research can not be carried out effectively. Therefore, the present invention optimizes the key technology of soil macroprotein extraction, hoping that the present invention can make a certain contribution in soil metaproteomics research. Contents of the invention [0004] The purpose of the pre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14C07K1/30
CPCC07K1/145C07K1/30
Inventor 李乃荟吴凤芝陈宇飞周新刚刘守伟潘凯
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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